EP4258036A1 - Drehscheibenmikroskopvorrichtung mit potentiell erhöhter bildauflösung - Google Patents

Drehscheibenmikroskopvorrichtung mit potentiell erhöhter bildauflösung Download PDF

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Publication number
EP4258036A1
EP4258036A1 EP23165590.3A EP23165590A EP4258036A1 EP 4258036 A1 EP4258036 A1 EP 4258036A1 EP 23165590 A EP23165590 A EP 23165590A EP 4258036 A1 EP4258036 A1 EP 4258036A1
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EP
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Prior art keywords
disk
pattern
emission
shaped body
excitation
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Pending
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EP23165590.3A
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English (en)
French (fr)
Inventor
Christian Seebacher
Rainer Uhl
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Till I D GmbH
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Till I D GmbH
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Pending legal-status Critical Current

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    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/06—Means for illuminating specimens
    • G02B21/08—Condensers
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
    • G02B27/58—Optics for apodization or superresolution; Optical synthetic aperture systems
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N21/6456—Spatial resolved fluorescence measurements; Imaging
    • G01N21/6458—Fluorescence microscopy
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0032—Optical details of illumination, e.g. light-sources, pinholes, beam splitters, slits, fibers
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0036—Scanning details, e.g. scanning stages
    • G02B21/0044—Scanning details, e.g. scanning stages moving apertures, e.g. Nipkow disks, rotating lens arrays
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/0052—Optical details of the image generation
    • G02B21/0076—Optical details of the image generation arrangements using fluorescence or luminescence
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/0004—Microscopes specially adapted for specific applications
    • G02B21/002—Scanning microscopes
    • G02B21/0024—Confocal scanning microscopes (CSOMs) or confocal "macroscopes"; Accessories which are not restricted to use with CSOMs, e.g. sample holders
    • G02B21/008—Details of detection or image processing, including general computer control
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/06—Means for illuminating specimens
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B21/00—Microscopes
    • G02B21/16—Microscopes adapted for ultraviolet illumination ; Fluorescence microscopes
    • G—PHYSICS
    • G02—OPTICS
    • G02B—OPTICAL ELEMENTS, SYSTEMS OR APPARATUS
    • G02B27/00—Optical systems or apparatus not provided for by any of the groups G02B1/00 - G02B26/00, G02B30/00
    • G02B27/10—Beam splitting or combining systems
    • G02B27/14—Beam splitting or combining systems operating by reflection only
    • G02B27/141—Beam splitting or combining systems operating by reflection only using dichroic mirrors
    • G—PHYSICS
    • G01—MEASURING; TESTING
    • G01N—INVESTIGATING OR ANALYSING MATERIALS BY DETERMINING THEIR CHEMICAL OR PHYSICAL PROPERTIES
    • G01N21/00—Investigating or analysing materials by the use of optical means, i.e. using sub-millimetre waves, infrared, visible or ultraviolet light
    • G01N21/62—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light
    • G01N21/63—Systems in which the material investigated is excited whereby it emits light or causes a change in wavelength of the incident light optically excited
    • G01N21/64—Fluorescence; Phosphorescence
    • G01N21/645—Specially adapted constructive features of fluorimeters
    • G01N2021/6463—Optics

Definitions

  • the present disclosure relates to the field of microscopy. More particularly, the present invention relates to devices for imaging fluorescent samples by employing multipoint excitation and the parallel detection of emission spots, which can be concentrated into smaller ones with enhanced numerical aperture.
  • confocal imaging relies on an array of moving pinholes on a rotating (spinning) Nipkov-disk for spatially filtering excitation an emission beamlets. Since such a series of pinholes scans an area in parallel, each pinhole is allowed to hover over a specific area for a longer amount of time, thereby reducing the excitation energy needed to illuminate a sample when compared to single point laser scanning microscopes.
  • a second rotating disk carrying a corresponding array of microlenses, is usually employed.
  • Patent publication DE 102015112960 B3 (corresponding to patent publication US10520713B2 ) discloses a spinning disk confocal microscope, which requires a single rotating disk, only. Suitable microoptical elements on the disk create a rotating excitation pattern in front of this disk, which is, using suitable optical elements, subsequently imaged into the sample plane of an objective lens.
  • the corresponding emission pattern is imaged by means of the same objective and spatially filtered using pinholes on the very same disk, whereby excitation and emission beam are separated respectively combined by dichroic filter-elements and the path-lengths of excitation and emission beam is adjusted by means of a suitable path-length difference-compensation so as to make the plane of the excitation pattern to coincide with the focal-plane of the emission pattern in the pinhole-plane.
  • Azuma & Kei enhance the numerical aperture of the emission spot-cones with the help of an additional set of microlenses on the pinhole-disk. They are positioned such that microlenses and pinholes are separated by an optical distance corresponding to 1/2 the focal length of the microlenses.
  • a pattern of microlenses spaced with a pitch of 500 ⁇ m and a camera-field of 15 x 15 mm is provided.
  • the present invention discloses one or more solutions to the aforementioned problems and disadvantages of the background art.
  • Other technical advantages of the present invention will be readily apparent to one skilled in the art from the following description and claims. Accordingly, the present invention is directed to devices for enabling observation of a fluorescent sample with a microscope according to the appended independent claims.
  • the present invention is directed to devices employing a single disk only.
  • the disk provides everything needed for spinning disk imaging with a desired degree of super-resolution, i.e., microoptical elements or microlenses for spot-generation, optionally microoptical elements or microlenses for condensing emission-spots, and - also optionally - additional pinholes for spatial filtering of the emission light, but not affecting the excitation light.
  • the microoptical elements may be in form of convex lenses or in form of concave lenses, preferably microlenses. Any other form of microoptical elements having adequate optical characteristics may be used as well. Further preferred embodiments are disclosed in the corresponding dependent claims.
  • the present invention is directed to an embodiment of a system for enabling observation of a fluorescent sample with a microscope, the device comprising:
  • the device may further comprise a second plurality of microoptical elements located on the disced shaped body at a second radial distance, r 1 , from the centre of the disk-shaped body, the microoptical elements exhibiting a second focal length, f 41 , and being located to form a pattern corresponding to the emission spot pattern.
  • the focal-length f 41 may assume values between infinity, in which case the NA of beamlets and hence the resolution remains unchanged, and a finite value, which leads to a doubling of the NA and provides maximal resolution-enhancement.
  • the at least partially light transmitting area may be configured as plurality of confocal pinholes, wherein the pinholes are located such that they provide spatial filtering for the emission spot pattern.
  • the present invention is directed to an alternative embodiment of a device for enabling observation of a fluorescent sample with a microscope, the device comprising:
  • the devices may further comprise a plurality of pinholes, each pinhole of the plurality of pinholes corresponding to and matching with the pattern a microoptical element of the plurality of microoptical elements being part of the emission pathway.
  • the plurality of pinholes may be arranged on an additional layer on the disk-shaped body, the additional layer being selectively transmitting the excitation light wavelengths but reflecting - outside the pinhole-openings - the emission wavelengths, thus constituting a spatial filter for emission light.
  • the pinholes may be etched into a dichroic layer, which may be located on the disked-shaped body itself, or it may be fixedly attached to the disked-shaped body.
  • the plurality of pinholes may be arranged on a second disk-shaped body.
  • the second disk-shaped body may be made of a material selectively transmits excitation light and constitutes a spatial filter for emission light.
  • the pinholes may be etched into the material being selectively transmission for excitation light and constitutes a spatial filter for emission light.
  • the pinholes may be arranged on a layer being selectively transmission for excitation light and constitutes a spatial filter for emission light, the layer may be positioned on the second disk-shaped body.
  • the excitation and emission beam are combined in an infinity optical space of the projection system relaying the emission spot-pattern to the camera. This is achieved by means of a dichroic element or pinhole in a reflecting element, which reflects > 99% of the emission light whereas it transmits a laser-spot, which is transformed into the collimated excitation beam illuminating the spot-forming plurality of microlenses.
  • the emission spot pattern may be imaged onto a detector through a projective lens system, forming the desired image when the first disk-shaped body, and, if present, the second disk-shaped body is rotated.
  • the device may further comprise a third dichroic element positioned in an infinity space in the projective lens system.
  • the emission spot pattern may be imaged onto a detector through a projective lens system, forming the desired image when the first disk-shaped body is rotated.
  • the device may further comprise a third reflective element positioned in an infinity space in the projective lens system.
  • the third reflective element may further comprises a hole, and the excitation beam may be directed through said hole before it is collimated by the projective lens system.
  • the hole may be positioned in the backfocal-plane of the third reflective element.
  • the projective lens system may comprise a first telecentric lens and a second telecentric lens.
  • the present invention extends the teaching of the prior art so as to achieve a resolution enhancement by using a single disk, only, for creating the rotating excitation pattern and for condensing the resulting emission spots into smaller ones with increased NA, which are subsequently imaged onto a camera-chip so as to form a resolution-enhanced image.
  • This "spot-condensation" is achieved with the help of an array of microoptical elements, preferably a microlens-array, arranged on the disk-shaped body, such as a rotating disk, in a pattern precisely matching the emission spot-pattern and being perfectly aligned with it.
  • the spot-condensing microlens-array is located on the very same rotating disk, which has generated the excitation pattern, and in one embodiment of the invention it even employs the same microlenses for excitation-pattern generation and for tighter focussing of the emission spot-pattern. In another embodiment, a set of different microlenses are used for excitation-pattern generation and for tighter focussing of the emission spot-pattern.
  • the microoptical elements may be in form of convex lenses and/or concave lenses.
  • the degree of focal-spot condensation obeys the Lagrange-Invariant principle, i.e. it increases the numerical aperture (NA) of the individual spot-cones accordingly and can maximally be adjusted to yield a twofold condensation.
  • NA numerical aperture
  • adjustment of the degree of resolution enhancement is achieved by tuning the path-length difference between the excitation- and emission-beam so as to yield the desired spot-condensation.
  • FIG. 1a showing a schematic illustration of a first embodiment of the present invention relying on a single disk-shaped body 32 rotatable around a central axis.
  • the disk-shaped body 32 contains a pattern of a first plurality microoptical elements, preferably microlenses 31 with a focal-plane 35.
  • Collimated excitation light 33 passes the first plurality of microoptical elements 31, thereby creating a spot-pattern 34 for excitation in front of the disk in the focal plane 35 of the first plurality of microoptical elements 31.
  • the resulting excitation beam 50 elicits an emission-pattern in the sample 9.
  • the emission beam 51 on its way back to the disk-shaped body 32, is separated from the excitation beam-path 50 by a single dichroic element 39. Between this dichroic element 39 and the disk 32 the two beams 50 respectively 51 propagate on different courses, use tube-lenses 36 respectively 37 of different focal lengths, f 36 respectively f 37 , whereby the number of reflections each encounters before reaching the disk 32 must be odd for both, even for both, or even. Accordingly, in the schematic the emission beam 51 reaches the disk with no reflections, whereas the excitation beam 50 is reflected by the dichroic element 39 and the reflective element, preferably a mirror 38 on its way to the microscope objective 40. Emission beam 51 and excitation beam 50 intersect the disk at different radii, r 1 and r 2 .
  • excitation takes the longer "tour", needs a correspondingly longer focal length (f 36 > f 37 ) for its tube-lens 36 and must originate from a microoptical element-pattern that is positioned closer to the outer rim of the disk, at r 2 , than the radius r 1 , at which the emission beam reaches the disk.
  • a matching pattern of a second plurality of microoptical elements, 41, preferably microlenses, scaled r 2 : r 1 with respect to the pattern of first plurality of microoptical elements 31, provides the concentration of the spots into smaller spots as needed for super-resolution.
  • Resolution enhancement is maximal if the focal plane of the tube-lens 37 in the emission beam-path is adjusted such relative to the microoptical elements that these microoptical elements 41 produce a twofold increase in NA.
  • the resulting image at a distance of 0.5*f 41 behind the second plurality of microoptical elements 41 is then projected onto a detector, such as a camera, using a suitable projection assembly.
  • the path-length can be adjusted to yield maximal resolution enhancement, but any NA-enhancement between 1x and 2x can be chosen at will in order to fine-tune the desired resolution enhancement.
  • the advantage of the embodiment depicted in figure 1a is that it merely needs a single dichroic element, and given this element is positioned in the infinity optical space, the dichroic layer can even be applied to a thick glass-substrate in order not to compromise dichroic flatness while at the same time avoiding spherical aberrations. Moreover, aligning the position of two beams 50, 51 relative to each other is more straightforward, but the tube lenses 36, 37 must exhibit equal distortion and a precisely adjustable r 1 /r 2 ratio.
  • out-of-focus rejection can be achieved by placing a matching pinhole layer 44, aligned with the second plurality of microoptical elements 41, at a distance required to afford the desired resolution enhancement.
  • the focal length of the microoptical elements 41 relative to the glass-thickness is chosen such that the desired resolution enhancement is achieved.
  • the focal length of the microoptical elements 41 is chosen such that the NA is increased twofold. A larger focal length, yielding less than a twofold NA-increase, reduces the degree of resolution enhancement.
  • microoptical elements 41 as shown in figure 1d , that is replacing them with flat surfaces at r 1 (in effect, by increasing their focal length to infinity), thereby forming an at least partially light transmitting area 48, no resolution enhancement is achieved.
  • the invention teaches a spinning disk-system that is perfectly functional for lower magnification objectives with large pupil which are poorly served by systems employing more than one disk for generating the excitation pattern and spatially filtering the emission pattern.
  • Figures 2a and 2b show an alternative embodiment according to the present invention, in which a suitable pattern of microoptical elements, such as microlenses 1 with focal-length f 1 , is applied to a rotating disk-shaped body 2 and is illuminated with collimated excitation light 3 from its rear side ( fig. 1a ).
  • the resulting excitation spot-pattern 4 in front of the disk, in the focal-plane 5 of the microoptical elements 1, is imaged, using a set of suitable optics elements 6, into a sample-plane 7 of a microscope, where it forms an excitation pattern 8 in a sample 9.
  • the beam between disk and sample is - in the following - called excitation image-beam 10.
  • the emission beam 11 On its reverse path ( fig. 2b ) the emission beam 11 is separated from the excitation beam-path 10 by means of a dichroic beamsplitter 12, and the beam-paths 10, 11 are reunited again by means of a second dichroic beamsplitter 13.
  • the emission beam takes a shortcut on its way back to the disk, thereby creating a path-length difference for the two beam-paths 10, 11, which can be adjusted such that the emission spot-pattern 14 does not fall into the same plane as the excitation spot pattern 5 in front of the disk 2, but significantly behind it.
  • the spots of the emission beam 11 pass the microoptical elements 1 backwards so as to form a 2x concentrated spot-pattern 15 at a distance of 0.5*f 1 behind the microoptical elements 1 in the disk 2.
  • the resulting image-beam 15, 43 comprising the NA-enhanced emission-pattern 15a, 42, is then imaged onto a camera 16, where it forms the desired super-resolution image when disk and hence pattern are rotated.
  • a pair of telecentric lenses 17, 18, the infinity space between these lenses is best suited for separating the excitation beam 3, 23 from the emission image beam 15, 43.
  • the tiny hole 20 To minimize the diameter of the tiny hole 20, it is positioned in the backfocal-plane of the first relay-lens 17, where the enhanced NA of emission beam 15 fills an area with 2x the diameter of the corresponding objective pupil (under the assumption of assumes equal focal lengths for relay-lens 17 and the tube lens of the microscope ( Fig. 2b ).
  • Excitation beam 3 33 has been called collimated, but its divergence-requirements are significantly relaxed compared to a diffraction-limited collimated beam.
  • a system featuring a plurality of microoptical elements to form multiple focussed spots tolerates a much wider beam-waist for beam 23 within hole 20 than an optical system where all light is focussed into a single spot.
  • a 100x 1.49 objective is used with a focal length of 200 mm for a tube-lens and relay-lens 17.
  • a diffraction limited spot in the focal plane 5 of the microoptical elements 1 has, at 488 nm, a FWHM diameter of 17.5 ⁇ m. If one accepts a geometrical extension of the spot in this plane of 10%, the beam diameter in the tiny through-hole 29 may be as wide as 95 ⁇ m.
  • the same relaxed divergence-requirements apply when the optical fiber is not fed by an incoherent light-source, but a multimode laser-source which employs speckle reduction techniques of prior art to generate a homogeneously radiating fiber exit area.
  • the optical scheme as shown in Figs 2a and 2b provides super-resolution, but no out-of-focus rejection, yet.
  • sectioning is obtained by placing a corresponding pinhole-pattern 21, matching the microoptical elements 1 used for excitation, at a suitable distance behind - as viewed from the microscope - the disk and aligning it with the microoptical elements 1.
  • the pinhole containing layer 22 must be selectively transparent for excitation light and constitute a spatial filter for emission wavelengths, only. This is achieved by etching the pinhole pattern 21 into a suitable dichroic layer 22.
  • the present invention is preferably used for fluorescence microscopy with one-photon excitation, or multi-photon excitation (mainly two photon excitations), so that the illumination light is excitation light and the light collected from the sample is fluorescence emission light; in one-photon excitation the emission light is of longer wavelength than the excitation light, and in multi-photon excitation the emission light is of shorter wavelength than the excitation light.
  • one-photon excitation the emission light is of longer wavelength than the excitation light
  • multi-photon excitation the emission light is of shorter wavelength than the excitation light.
  • references in the appended claims to an method or device or a component of an device or system being adapted to, arranged to, capable of, configured to, enabled to, operable to, or operative to perform a particular function encompasses that apparatus, system, component, whether or not it or that particular function is activated, turned on, or unlocked, as long as that apparatus, system, or component is so adapted, arranged, capable, configured, enabled, operable, or operative. Additionally, although this disclosure describes or illustrates particular embodiments as providing particular advantages, particular embodiments may provide none, some, or all of these advantages.

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  • Physics & Mathematics (AREA)
  • General Physics & Mathematics (AREA)
  • Optics & Photonics (AREA)
  • Chemical & Material Sciences (AREA)
  • Analytical Chemistry (AREA)
  • Health & Medical Sciences (AREA)
  • Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
  • Life Sciences & Earth Sciences (AREA)
  • Biochemistry (AREA)
  • General Health & Medical Sciences (AREA)
  • Immunology (AREA)
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  • Microscoopes, Condenser (AREA)
EP23165590.3A 2022-04-07 2023-03-30 Drehscheibenmikroskopvorrichtung mit potentiell erhöhter bildauflösung Pending EP4258036A1 (de)

Applications Claiming Priority (1)

Application Number Priority Date Filing Date Title
DE102022108448.5A DE102022108448B3 (de) 2022-04-07 2022-04-07 Superauflösende Mikroskopvorrichtung mit rotierender Scheibe

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EP4258036A1 true EP4258036A1 (de) 2023-10-11

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Citations (8)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
JPH09133870A (ja) 1995-11-10 1997-05-20 Yokogawa Electric Corp 共焦点光スキャナ
EP0753779B1 (de) 1995-07-13 2003-09-10 Yokogawa Electric Corporation Konfokales Mikroskop
DE102007009551B3 (de) 2007-02-27 2008-08-21 Ludwig-Maximilian-Universität Vorrichtung für die konfokale Beleuchtung einer Probe
DE102013001238A1 (de) * 2013-01-25 2014-07-31 Carl Zeiss Microscopy Gmbh Lichtmikroskop und Mikroskopieverfahren
DE102015112960B3 (de) 2015-08-06 2016-10-20 Till I.D. Gmbh Vorrichtung für die konfokale Beleuchtung einer Probe
CA2978123A1 (en) * 2017-09-05 2019-03-05 Peter Vokhmin Real time multichannel sted microscopy system
US20190187447A1 (en) * 2017-12-20 2019-06-20 Olympus Corporation Disk scanning microscope system and computer-readable recording medium
US10352860B2 (en) * 2014-04-24 2019-07-16 Bruker Nano, Inc. Super resolution microscopy

Family Cites Families (1)

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Publication number Priority date Publication date Assignee Title
JP2004138819A (ja) 2002-10-17 2004-05-13 Olympus Corp 蛍光共焦点顕微鏡

Patent Citations (9)

* Cited by examiner, † Cited by third party
Publication number Priority date Publication date Assignee Title
EP0753779B1 (de) 1995-07-13 2003-09-10 Yokogawa Electric Corporation Konfokales Mikroskop
JPH09133870A (ja) 1995-11-10 1997-05-20 Yokogawa Electric Corp 共焦点光スキャナ
DE102007009551B3 (de) 2007-02-27 2008-08-21 Ludwig-Maximilian-Universität Vorrichtung für die konfokale Beleuchtung einer Probe
DE102013001238A1 (de) * 2013-01-25 2014-07-31 Carl Zeiss Microscopy Gmbh Lichtmikroskop und Mikroskopieverfahren
US10352860B2 (en) * 2014-04-24 2019-07-16 Bruker Nano, Inc. Super resolution microscopy
DE102015112960B3 (de) 2015-08-06 2016-10-20 Till I.D. Gmbh Vorrichtung für die konfokale Beleuchtung einer Probe
US10520713B2 (en) 2015-08-06 2019-12-31 Till I.D. Gmbh System for confocal illumination of a sample
CA2978123A1 (en) * 2017-09-05 2019-03-05 Peter Vokhmin Real time multichannel sted microscopy system
US20190187447A1 (en) * 2017-12-20 2019-06-20 Olympus Corporation Disk scanning microscope system and computer-readable recording medium

Non-Patent Citations (1)

* Cited by examiner, † Cited by third party
Title
AZUMAKEI: "Optics Express", vol. 23, 2015, article "Super-resolution spinning-disk confocal microscopy using optical photon re-assignment", pages: 15004

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DE102022108448B3 (de) 2023-05-04

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